Self-assembled nano-Ag/Au@Au film composite SERS substrates show high uniformity and high enhancement factor for creatinine detection

Serum creatinine is a key biomarker for the diagnosis and monitoring of kidney disease. Rapid and sensitive creatinine detection is thus important. Here, we propose a high-performance nano-Ag/Au@Au film composite SERS substrate for the rapid detection of creatinine in human serum. Au nanoparticles (AuNPs) and Ag nanoparticles (AgNPs) with uniform particle size were synthesized by a chemical reduction method, and the nano-Ag/Au@Au film composite SERS substrate was successfully prepared via a consecutive layer-on-layer deposition using an optimized liquid-liquid interface self-assembly method. The finite element simulation analysis showed that due to the multi-dimensional plasmonic coupling effect formed between the AuNPs, AgNPs, and the Au film, the intensity of the local electromagnetic field was greatly improved, and a very high enhancement factor (EF) was obtained. Experimental results showed that the limit of detection (LOD) of this composite SERS substrate for rhodamine 6G (R6G) molecules was as low as 1 × 10^-13M, and the Raman EF was 15.7 and 2.9 times that of the AuNP and AgNP monolayer substrates respectively. The results of different batch tests and SERS mapping showed that the relative standard deviations of the Raman intensity of R6G at 612 cm^-1were 12.5% and 11.7%, respectively. Finally, we used the SERS substrate for the label-free detection of human serum creatinine. The results showed that the LOD of this SERS substrate for serum creatinine was 5 × 10^-6M with a linear correlation coefficient of 0.96. In conclusion, the SERS substrate has high sensitivity, good uniformity, simple preparation, and has important developmental potential for the rapid detection and application of disease biomarkers.